197 results about "Cell factory" patented technology

The invention relates to the field of biological engineering and technology, in particular to an enzyme for synthesizing glabridin, a biological synthesis method of glabridin and a bacterial strain. The invention discloses a series of glabridin synthesizing enzymes (phenylalanine ammonialyase, cinnamic acid-4-hydrogenase, tyrosine ammonialyase, p-coumaric acid coenzyme A ligase, chalcone synthase, chalcone reductase, chalcone isomerase, isoflavone synthase, 2-hydroxyisoflavone dehydratase, 4 '-oxymethyltransferase, 4'-hydroxymethyltransferase and 4 '-hydroxymethyltransferase) derived from glycyrrhiza glabra for the first time. ) can be used in the preparation of a compound (e.g., isoflavone 2 '-hydrogenase, isoflavone reductase, Vissisoketoreductase, isoprenyltransferase, pterocarpin reductase, oxidative cyclase, demethylase, cytochrome P450 reductase, and the like). The de novo synthesis of the glabridin in a microbial cell factory is realized for the first time, and the application prospect of the glabridin biological manufacturing industry is greatly promoted.

The invention discloses a rare ginsenoside dammadienol synthetase and an application of the rare ginsenoside dammadienol synthetase in preparation of dammadienol. The dammadienol synthetase is identified for the first time in the research, a metabolic network of a saccharomyces cerevisiae host cell is modified through a metabolic engineering strategy, genes which do not exist in the host cell are introduced into an engineering strain for heterologous expression, the host cell is changed from nothing to existence, and the dammadienol engineering strain is constructed. In addition, through homologous modeling, key catalytic pocket searching and key enzyme gene site-directed mutagenesis modification, enzyme catalysis efficiency is improved, efficient synthesis of dammadienol is achieved, the problem that natural sources of the rare ginsenoside are insufficient is solved, green synthesis and sustainable utilization of the rare ginsenoside are promoted, and the method is suitable for industrial production. And a foundation is laid for subsequent construction of rare ginsenoside cell factories.

The invention discloses a novel biosynthesis path for producing 3 '-phosphoadenosine-5'-phosphosulfuric acid and application, and belongs to the technical field of bioengineering. According to the invention, through recombinant expression and purification of ribose kinase, ribose phosphate pyrophosphate kinase, ribose phosphate transferase, adenine thiotransferase, adenosine phosphate sulfate kinase and polyphosphate kinase 2, a six-enzyme cascade reaction taking ribose and adenosine as substrates in vitro is constructed, the molar conversion rate reaches 22.85%, and the escherichia coli and bacillus subtilis PAPS cell factory can be used for preparing the recombinant ribose phosphate pyrophosphate kinase, the ribose phosphate transferase, the adenine thiotransferase, the adenosine phosphate sulfate kinase and the polyphosphate kinase 2. The yield of PAPS in a 5-L fermentation tank reaches 7.60 g / L and 5.03 g / L respectively. The PAPS synthesis route provided by the invention provides an extensible, economical and efficient solution for the production of sulfated compounds.

The invention relates to recombinant escherichia coli and a method for synthesizing sialic acid lactose-N-tetrasaccharide c, and belongs to the technical field of microbial metabolism engineering. According to the research, engineering escherichia coli BAZ for producing LNTri II is taken as an initial strain, two key glycosyl transferases, namely beta 1, 4-galactosyl transferase and alpha 2, 6-sialyltransferase, and three CMP-Neu5Ac pathway enzymes, namely NeuC, NeuB and NeuA, are introduced, and related competitive genes are knocked out; under the conditions of shake-flask culture and fed batch culture, the LST c yield of 1.718 g / L and the LST c yield of 9.745 g / L are achieved through the engineering bacterium respectively, and the huge potential of a microbial cell factory in the aspect of efficient production of complex breast milk oligosaccharides is shown.

The invention discloses a novel plasmid vector capable of increasing CHO foreign protein expression quantity and construction and application thereof, and belongs to the technical field of gene engineering. The plasmid vector pCHO6GS belongs to a brand-new plasmid system, and the expression quantity of a common monoclonal antibody and recombinant protein (such as H5HA trimer) difficult to express in CHO can be increased by at least more than one time, so that the expression quantity of foreign protein is increased beyond expectation through the novel plasmid vector; and the method also plays an extremely important role in reducing cost and improving efficiency of various protein products taking CHO as a cell factory.

The invention discloses a yeast engineering bacterium for producing apigenin and application thereof. The yeast engineering bacterium comprises a yeast genome, a whole-cell catalytic synthesis apigenin module, a heterologous apigenin biosynthesis path and an NADPH supply module, the whole-cell catalytic synthesis apigenin module comprises an FNSI coding gene and an FNSII coding gene; the heterologous apigenin biosynthesis path comprises a TAL coding gene, a 4CL coding gene, a CHS coding gene, a CHI coding gene and an FNSII coding gene; the NADPH supply module comprises a G6PD coding gene and a PGD coding gene; the method disclosed by the invention has the characteristics of greenness and safety, and lays a foundation for synthesizing apigenin in a yeast cell factory.

The invention discloses a bifunctional flavin adenine dinucleotide synthetase mutant and an application of the bifunctional flavin adenine dinucleotide synthetase mutant in FAD synthesis. The mutant disclosed by the invention is RFK / FADSC18R or RFK / FADSC18R-R46H, and the structural formula of the mutant is as shown in the specification. The amino acid sequence of the RFK / FADSC18R is as shown in SEQ ID NO. 15, and the amino acid sequence of the RFK / FADSC18R-R46H is as shown in SEQ ID NO. 16. A genetically engineered bacterium carrying the bifunctional flavin adenine dinucleotide synthetase mutant gene plasmid is used for producing flavin adenine dinucleotide by using glucose as a substrate through a simple fermentation process under a shake flask aerobic condition, and the FAD yields of strains XF02 and XF03 containing mutant genes are 79.31 mg / l and 167.56 mg / l in sequence; and compared with a strain XF01 containing a non-mutated gene, the mutant gene is improved by 58.62% and 235.12% respectively. A microbial'cell factory 'is constructed through a metabolic engineering strategy, and the modified microorganisms can generate required natural products in microbial cells by utilizing low-price raw materials. The progress not only reduces the production cost of the FAD, but also improves the environmental sustainability of the production process of the FAD.

The invention belongs to the technical field of microbial engineering and genetic engineering, and discloses escherichia coli for producing histidine as well as a construction method and application thereof. The escherichia coli contains a growth dependent promoter PprpB of a methyl isocitrate lyase gene prpB and / or a stable phase dependent promoter PydgI of an ornithine reverse transport protein / lysine permease gene ydgI. Escherichia coli for producing histidine uses growth dependent promoter PprpB of methylisocitrate lyase gene prpB to regulate expression of glutamine phosphoribose pyrophosphate transamidase gene purF and uses ornithine reverse transport protein / lysine permease gene (ydgI) The stable-phase dependent promoter PydgI regulates the expression of the ATP phosphoribosyltransferase gene hisG, so that the energy supply of thalli in early-stage cell growth and the metabolic flux of L-histidine in later-stage thalli production can be realized, and the balanced distribution of the metabolic flux between the cell growth of a microbial cell factory and the production of L-histidine is effectively balanced.

The invention discloses a directed evolution strategy of type II sesquiterpene cyclase (SsDMS) derived from Streptomyces shedoensis based on alanine scanning, and an application of the type II sesquiterpene cyclase (SsDMS) derived from Streptomyces shedoensis based on alanine scanning. The invention relates to a plurality of SsDMS key mutant sites capable of catalytically generating a novel drive type sesquiterpenoid compound, which comprise substitution of sites 208, 248, 249, 497 and 505, and the SsDMS key mutant sites comprise substitution of sites 208, 248, 249, 497 and 505. The invention discloses an efficient escherichia coli cell factory applied to heterologous expression of an SsDMS mutant and efficient production of a novel drive type sesquiterpene compound, and further obtains the drive type sesquiterpene compound with a novel structure. According to the present invention, the efficient production of the drive type sesquiterpene compound by using the SsDMS and the mutant thereof is achieved, the modification strategy has wide applicability, the important enzymology resource is provided for the biosynthesis of the novel terpene compound, and the more choices are provided for the discovery of the drug lead compound.

The invention belongs to the technical field of rabies vaccines, and particularly relates to a purification method of a rabies virus inactivated vaccine, which comprises the following steps: step 1, carrying out passage by using a Walvax-2 cell strain through a cell factory method, and carrying out multiplication culture on the passage production cells in a microcarrier bioreactor; 2, cleaning the cells with a treating fluid, inoculating the cells with a rabies virus fixed strain, and preparing and culturing a rabies virus solution; step 3, carrying out ultrafiltration concentration on the rabies virus stock solution; step 4, purifying the rabies virus concentrated solution; step 5, inactivating the rabies virus; and 6, hydrolyzing to obtain the rabies vaccine stock solution. By adopting the composite Capto Core 700 column chromatography, impure proteins can be effectively removed, the recovery rate is high, the quality of the rabies vaccine obtained by adopting a mode of firstly purifying and then adopting beta-propiolactone inactivation is excellent, and the effect of the finally obtained human diploid cell rabies vaccine stock solution is good.

The invention belongs to the technical field of microbial metabolism engineering and industrial biology, and relates to an engineering bacterium for producing fulvic alcohol and a construction method and application thereof. The engineering bacteria capable of increasing the content of demethylated fulvic alcohol serving as a synthetic precursor are obtained by regulating and controlling the metabolic pathway synthetase of fulvic alcohol in a host strain, and then the methyltransferase of the engineering bacteria is optimized, so that the engineering bacteria capable of producing fulvic alcohol are obtained. Wherein the host strain is selected from any saccharomyces cerevisiae carrying a fulvic alcohol path. The yeast cell factory is constructed by coordinating and balancing a plurality of precursor module metabolic flows, and the heavy synthesis of the fulvic alcohol is realized for the first time.

The invention relates to the technical field of synthetic biology, in particular to a microbial cell factory for producing catechol. Pseudomonas putida without a pcaHG gene, a catA gene and a catA2 gene is used as a chassis bacterium, and exogenous AroY and / or kpdB genes are / is introduced into the chassis bacterium, or the expression of endogenous genes VanAB and PobA is improved. The lignin-derived monomer is efficiently converted into catechol through the strategy of cofactor regulation and key enzyme overexpression, the conversion rate reaches 98.5%, the highest yield is 14.1 mM, and compared with the previous 64% conversion rate, the conversion rate is greatly improved.

The present disclosure relates to a genetically modified host cell having improved production of sulfurol and / or sulfurol precursors, wherein the host cell expresses one or more heterologous thiaminase I, thiaminase II and / or phosphatase enzymes and / or one or more heterologous thiamine pathway enzymes converting thiamine, 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic phosphate acid (cTHZ-P), 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic acid (cTHZ) and / or THZ-P to sulfurol and / or 5-(2-hydroxyethyl)-4-methyl-1,3-thiazole-2-carboxylic acid (cTHZ), whereby the production of the sulfurol in the genetically modified host cell is improved compared to an unmodified parent host cell. Chemical methods for production of sulfurol are also provided.

The invention discloses a bacillus constitutive strong promoter and application thereof, and relates to the technical field of biology, and the nucleotide sequence of the promoter is SEQ ID NO: 2. The mRNA transcriptional level of the reporter gene is improved; the method is used for starting a target gene to efficiently express amylase in bacillus subtilis. The method is also used for starting a target gene to efficiently express alkaline protease in bacillus licheniformis. The novel constitutive promoter with enhanced transcriptional activity is developed by taking P43 as a starting template, and under the same host and culture conditions, the absolute level of the constitutive promoter for driving expression of a reporter gene or a target product is higher than that of the current P43 promoter, so that core power is provided for performance improvement of a bacillus cell factory.

The invention belongs to the field of synthetic biology, and particularly relates to a genetically engineered bacterium for producing O-succinyl-L-homoserine at high yield as well as a construction method and application of the genetically engineered bacterium. According to the invention, the large intestine probiotic Nissle 1917 is used as a chassis strain, and compared with other expression hosts, the large intestine probiotic Nissle 1917 can grow and breed more quickly and is safer. The expression of the gene thrB is regulated by using the self-regulation promoter PfliA, so that the synthesis route of threonine is dynamically regulated, the efficient utilization of a carbon source is realized, and the sugar acid conversion rate of OSH synthesis is improved; in addition, by introducing site mutation, feedback inhibition suffered by OSH synthesis key enzyme HST is relieved, the enzyme activity of consumption pathway key enzyme CGS is reduced, and the carbon flow is effectively driven to be redirected to a target product synthesis pathway. According to the microbial cell factory constructed by the invention, the production efficiency of OSH is remarkably improved, the fermentation period is shortened, the production cost is reduced, and the economic feasibility of industrial production is realized.

The invention provides a cell strain for producing engineered viroid particles, a method thereof and the engineered viroid particles, the cell strain is named as SZCV-Con28 and preserved in Guangdong Microbial Culture Collection Center on October 15, 2025, the preservation number is GDMCC No: 67101, and the preservation number is GDMCC No: 67101. A plurality of corresponding cell factories for producing the engineered viroid particles are established by cell lines obtained by introducing different target gRNA elements into the cell strain, so that the corresponding engineered viroid particles can be continuously produced in batches. According to the invention, large-scale preparation can be realized through cell expanding culture, the capacity limitation is overcome, the production process flow is simplified, and the consumption of production raw materials is reduced.

The invention discloses curcuma wenyujin sesquiterpene synthase CwTPS8 as well as a coding gene and application thereof, and belongs to the technical field of plant genetic engineering, the amino acid sequence of the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.1, the nucleotide sequence of the gene for coding the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.2, and the nucleotide sequence of the gene for coding the curcuma wenyujin sesquiterpene synthase CwTPS8 is shown as SEQ ID NO.1. The curcuma wenyujin sesquiterpene synthase CwTPS8 can be used for preparing beta-caryophyllene, alpha-humulene and other curcuma wenyujin sesquiterpenoids by taking farnesyl pyrophosphate FPP as a substrate, and the technology can be used for subsequently producing the sesquiterpenoids through a microbial cell factory. Meanwhile, a foundation is laid for heterologous synthesis and further development and utilization of sesquiterpenoids such as beta-caryophyllene and alpha-humulene.

The invention provides a method for controlling pests by utilizing intestinal symbiotic bacteria to express dsRNA (double-stranded ribonucleic acid) and application of the method. According to the invention, intestinal symbiotic bacteria are used as a cell factory, and interfering molecules (such as dsRNA) are continuously generated in intestinal tracts of pests to interfere the expression of target genes of the pests, so that the pests are effectively and continuously controlled.

This invention aims to create a spatially isolated local reaction microenvironment for Rubisco by introducing a complete β-carboxysome microcompartment system. This design goal avoids oxygen inhibition, laying the foundation for ultimately achieving efficient and specific CO2 fixation. It provides a novel approach to overcome the fundamental problem of low efficiency of carbon-fixing enzymes in cytosols due to intense oxygen competition. By introducing the complete carboxysome system as a module, engineered yeast strains with high carboxylase activity and significantly improved ethanol production were obtained, providing a solid foundation for building next-generation yeast cell factories that produce high-yield ethanol using CO2 as a raw material.

This invention relates to the field of synthetic biology, and more particularly to a microbial cell factory for producing catechols. The invention uses *Pseudomonas putida* strains with the pcaHG, catA, and catA2 genes knocked out as substrate bacteria, and introduces exogenous AroY and / or kpdB genes, or increases the expression of endogenous genes VanAB and PobA. Through a strategy of cofactor regulation and key enzyme overexpression, lignin-derived monomers are efficiently converted into catechols, achieving a conversion rate of 98.5% and a maximum yield of 14.1 mM, a significant improvement compared to the previous conversion rate of 64%.

The utility model provides a liquid discharge water tank device suitable for a cell factory, and particularly provides a liquid discharge water tank device which sequentially comprises a tank body part, a flow guide bottom plate, a liquid discharge port, a supporting base and a connecting pipeline from top to bottom, the flow guide bottom plate and the liquid discharge port are arranged at the bottom of the tank body, and the connecting part of the flow guide bottom plate and the liquid discharge port is low in height. The part far away from the liquid outlet is high, the liquid outlet is connected with the connecting pipeline, and a clamping groove is formed in the tank body part and used for fixing a cell factory, so that the liquid outlet of the cell factory is suspended above the liquid discharging water tank device. When the device is applied, waste liquid in the cell factory is directly discharged from the liquid outlet of the cell factory without a solution remaining area, so that the defect that waste liquid is easy to remain at a pipeline connecting port in the prior art is overcome, the part in contact with the waste liquid is detachable, the sterilization is easy, the operation is convenient, the liquid discharging speed is high, and waste liquid accumulation is hardly generated; when the device is used for discharging waste liquid in a cell factory, residual impurities on the cell surface are lower.

The invention relates to a high-titer influenza virus seed as well as a preparation method and application thereof, and the preparation method comprises the following steps: S1, obtaining a virus, inoculating the virus into a cell factory, and culturing to obtain a cell supernatant; and S2, carrying out microfiltration on the cell supernatant to remove cell debris in the cell supernatant, carrying out ultrafiltration concentration, and carrying out degerming filtration to obtain the high-titer influenza virus seed. According to the technical scheme, the cell factory is adopted for culturing the virus, the virus is inoculated to the ten-layer cell factory, after culturing, the cell supernatant in the cell factory is harvested, microfiltration is carried out, and the cell debris is removed, so that compared with a conventional method for removing the cell debris through centrifugation, microfiltration needs shorter time, the volume of the supernatant which can be treated at a time is larger, and the cell debris can be removed at a time. The sterile risk is lower; and concentrating the supernate, and sterilizing and filtering after concentration. The virus titer of the virus seed subjected to the preparation process can be increased by 5-10 times, and the impurity content is lower.

A simpler and lower-cost method and associated hardware are described for recovering biopharmaceutical products from cell factories in higher yields by a liquid nitrogen-based pre-treatment of cell factories followed by gentler disintegration of cellular factories by tunable, pressurizing and de-pressurizing cycle with gaseous nitrogen. The cell factories are heterologous expression systems and could be based on plant cells, mammalian cells, algal cells, yeast cells or even bacterial cells. The pre-treatment step with liquid nitrogen softens the tougher cell wall structures thus enabling cells to be disrupted relatively easily during the gaseous nitrogen-based pressurizing / depressurizing cycle. This invention can enhance the yield of biopharmaceutical products from expression systems and reduce the overall cycle-time and further down-stream processing and polishing cost of biopharmaceutical products.

The invention provides a method for dynamically regulating and controlling the copy number of plasmids in saccharomyces cerevisiae based on RNA (Ribonucleic Acid) interference. According to the method, an orthogonal RNAi system is reconstructed, saccharomyces cerevisiae is used as a synthetic biological chassis with a dynamic programmable plasmid copy number, a heterologous RNAi mechanism from Ckstri saccharomyces cerevisiae is integrated, sequence-specific siRNA of a targeted plasmid coding selective marker is designed, a chemically induced gene dose control platform is established, and the gene dose control platform is used for controlling the gene dose. The dynamic regulation and control of the plasmid copy number are realized, so that the gene expression dosage is optimized, and the biosynthesis efficiency is improved. RNAi-mediated copy number regulation is applied to a carotenoid biosynthesis pathway, and compared with a static plasmid system, the lycopene titer is increased by 18.6 times. The method further enriches an anabolism regulation tool set, has huge potential in the aspect of improving the production performance of a microbial cell factory, and provides a new thought for metabolic engineering optimization in the field of synthetic biology.

The invention relates to polymerized glutamate sugar ester as well as a biological fermentation preparation method and application thereof, and belongs to the technical field of biological fermentation. According to the scheme, amino acid and functional oligosaccharide are taken as substrates, and efficient directional assembly of high-molecular polymerized glutamate sugar ester is realized by designing an integrated cell factory biosynthesis technology. The invention provides a brand-new polyglutamic acid sugar ester compound and a biological fermentation method of the polyglutamic acid sugar ester, the biological fermentation method is suitable for industrial continuous production, and the prepared polyglutamic acid sugar ester is narrower in molecular weight distribution and better in product uniformity.

The application discloses a kind of biological immobilized enzyme based on yeast surface display technology and the application of synthesis 2-phenylethanol, using yeast surface display technology, for the first time with Saccharomyces cerevisiae surface protein a-lectin as anchor protein, the key enzyme (transaminase, decarboxylase, ethanol dehydrogenase) involved in 2-PE producing Ehrlich pathway in Saccharomyces cerevisiae is separately or jointly carried out biological fixation, enhance substrate channel effect and proximity effect, in vitro one-step catalysis L-phenylalanine synthesis 2-PE, provide yield.The method for fixing multiple enzymes of Ehrlich pathway in the application can be used to reconfigure the biosynthesis pathway of other natural products in vitro, to provide feasible scheme for the rational design of cell factory.

The application belongs to the technical field of microbial sewage treatment, and particularly relates to an iron-carbon quantum dot / electrically active non-oxygen-producing photosynthetic bacteria / carrier hybrid cell factory and application thereof in sewage resourceization or carbon emission reduction; the iron-carbon quantum dot in the hybrid cell factory provided by the application promotes the non-oxygen-producing purple photosynthetic bacteria to utilize more light energy, and generates photo-generated electrons as an extracellular efficient electron donor source, and the photosensitive active substances in the extracellular polymeric substance of the non-oxygen-producing purple photosynthetic bacteria can promote the formation of a continuous cycle of trivalent iron and divalent iron, realize mutual cooperation, promote sewage resourceization and carbon emission reduction, and synthesize high-value-added products; and in addition to facilitating the recovery of biomass, the transparent conductive glass electrode as the carrier also ensures that the non-oxygen-producing purple photosynthetic bacteria stably and efficiently utilize light energy; therefore, the hybrid cell factory provided by the application is a high-efficiency microbial sewage treatment technology, and solves the technical problem of lack of high-efficiency microbial sewage treatment technology in the prior art.

The present disclosure relates to genetically modified host cells producing prostaglandins, to genetic constructs for expression of such mutants; to cultures of the genetically modified host cells and its use to produce the said products; to fermentation liquids comprising the said products resulting from such production; to compositions comprising the fermentation liquid; to pharmaceutical preparations made from such compositions and to the uses of such compositions and preparations.

The present invention relates to a method for constructing an engineered cyanobacteria that tolerates high temperature and high light, including the step of increasing the expression amount of shikimate kinase in the cyanobacteria; and also relates to an engineered cyanobacteria that grows rapidly under high temperature and high light conditions constructed on the basis of the engineered cyanobacteria that tolerates high temperature and high light and a construction method thereof. The engineered cyanobacteria constructed by the present invention can grow rapidly and stably under high temperature and high light conditions that the wild type cannot tolerate, and solves the problem of decreased or unstable target product yield caused by high temperature and high light stress in outdoor large-scale cultivation. In addition, whether under normal temperature and normal light conditions or under high temperature and high light stress conditions, the engineered cyanobacteria of the present invention has a significantly higher growth rate than its wild type, can accumulate more biomass, synthesize more glycogen, and can accumulate more protein under high temperature and high light conditions, so the chassis bacteria as a "cell factory" have the potential to produce target products more efficiently.

The invention discloses a double-fucosyl transferase complex and application thereof in synthesis of difucosyl lactose, and belongs to the technical field of biology. In order to solve the problem of high residual quantity of a by-product 2 '-FL in the existing difucosyl lactose (DFL) cell factory, the invention constructs a novel difucosyl transferase complex, random mutation and screening are performed on a key element connecting peptide in the complex, and the catalytic effect of an enzyme is optimized. Through the strategy, the constructed recombinant strain can control the residual quantity of a byproduct 2 '-FL while efficiently synthesizing the DFL, so that the subsequent separation and purification cost is remarkably reduced, the production efficiency is improved, and important data support is provided for industrial large-scale production of the DFL.